DOCSLIB.ORG

Macrocyclization of an All-D Linear Peptide Improves Target Affinity and Imparts Cellular Activity: a Novel Stapled −Helical Peptide Modality

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Macrocyclization of an All-D Linear Peptide Improves Target Affinity and Imparts Cellular Activity: a Novel Stapled −Helical Peptide Modality bioRxiv preprint doi: https://doi.org/10.1101/767673; this version posted September 12, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Macrocyclization of an all-D linear peptide improves target affinity and imparts cellular activity: A novel stapled −helical peptide modality Srinivasaraghavan Kannana*, Pietro G. A. Aronicaa, Simon Ngb, Dawn Theanb, Yuri Frosib, Sharon Cheeb, Jiang Shiminb, Tsz Ying Yuenc, Ahmad Sadruddind, Hung Yi Kristal Kaand, Arun Chandramohand, Jin Huei Wonga, Yaw Sing Tana, Fernando J. Ferrerb, Prakash Arumugama, Yi Hane, Shiying Chene, Christopher J. Brownb, Charles W. Johannesc, Brian Henryd, David P. Laneb, Tomi K. Sawyerf, Chandra S. Vermaa,g,h*., Anthony W. Partridged* aBioinformatics Institute, Agency for Science, Technology and Research (A*STAR), 30 Biopolis Street, #07-01 Matrix, Singapore 138671. bp53 Laboratory, Agency for Science, Technology and Research (A*STAR), 8A Biomedical Grove, #06-04/05, Neuros/Immunos, Singapore 138648. cInstitute of Chemical & Engineering Science, Agency for Science, Technology and Research (A*STAR), 8 Biomedical Grove, #07, Neuros Building, Singapore 138665. dMSD International, Translation Medicine Research Centre, Singapore. eMerck & Co., Inc., Kenilworth, New Jersey, USA. fMerck & Co., Inc., Boston, Massachusetts, USA. gSchool of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive,Singapore 637551. hDepartment of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543. *Corresponding authors: Srinivasaraghavan Kannan, Bioinformatics Institute (A*STAR), 30, Biopolis Street, 07-01 Matrix, Singapore 138671, Singapore, E-mail: [email protected] star.edu.sg; Tel: +65 6478 8353; Fax: +65 6478 9048. Chandra S. Verma, Bioinformatics Institute (A*STAR), 30, Biopolis Street, 07-01 Matrix, Singapore 138671, Singapore, E-mail: [email protected]; Tel: +65 6478 8273; Fax: +65 6478 9048. Anthony Partridge, MSD International, Translation Medicine Research Centre, 8 Biomedical Grove, #04-01/05 Neuros Building,Singapore. 138665, Singapore, E-mail: [email protected]. 1 bioRxiv preprint doi: https://doi.org/10.1101/767673; this version posted September 12, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract Peptide-based inhibitors hold great potential for targeted modulation of intracellular protein- protein interactions (PPIs) by leveraging vast chemical space relative to primary structure via sequence diversity as well as conformationally through varying secondary and tertiary structures. However, the development of peptide therapeutics has been hindered because of their limited conformational stability, proteolytic sensitivity and cell permeability. Several contemporary peptide design strategies address these issues to varying degrees. Strategic macrocyclization through optimally placed chemical braces such as olefinic hydrocarbon crosslinks, commonly referred to as staples, may address these issues by i) restricting conformational freedom to improve target affinities, ii) improving proteolytic resistance, and iii) enhancing cell permeability. Conversely, molecules constructed entirely from D-amino acids are hyper-resistant to proteolytic cleavage, but generally lack conformational stability and membrane permeability. Since neither approach is a complete solution, we have combined these strategies to identify the first examples of all-D α-helical stapled and stitched peptides. As a template, we used a recently reported all D-linear peptide that is a potent inhibitor of the p53-Mdm2 interaction, but is devoid of cellular activity. To design both stapled and stitched all-D-peptide analogues, we used computational modelling to predict optimal staple placement. The resultant novel macrocyclic all D-peptide was determined to exhibit increased α-helicity, improved target binding, complete proteolytic stability and, most notably, cellular activity. 2 bioRxiv preprint doi: https://doi.org/10.1101/767673; this version posted September 12, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Introduction Protein-protein interactions (PPIs) are central to most biological processes and are often dysregulated in disease [1, 2] . Therefore, PPIs are attractive therapeutic targets for novel drug discovery. However, in contrast to the deep protein cavities that typically accommodate small molecules, PPI surfaces are generally large and flat, and this has contributed to the limited success in developing small molecule inhibitors for PPI targets [3]. The realization that ~40% of all PPIs are mediated by relatively short peptide motifs gave rise to the possibility of developing peptide-based inhibitors that would compete orthosterically for the interface between ligand–target cognate partners [4]. When taken out of the protein ligand context and synthesized, such peptides may often be unstructured and intrinsically disordered, yet able to achieve their biologically-relevant conformation upon protein target binding [4]. However, for intracellular targets, the peptide modality may be challenging due to proteolytic sensitivity, low conformational stability (yielding weak affinities and off-target effects), and poor cell permeability (further limiting engagement of intracellular targets and/or oral bioavailability) [5-11]. To address these issues, several strategies have been pursued, including macrocyclization and modifications of the peptide backbone to yield molecules with improved biological activities and pharmacokinetic properties as well as constraining the peptide into its biologically-relevant conformation that binds its target [5-13]. First, by constraining the peptides toward their bound conformations, entropic penalties upon binding are reduced, thus improving binding constants as well as potentially decreasing the opportunity for unwanted off-target effects. Secondly, macrocyclization may confer varying degrees of proteolytic resistance by modifying key backbone and/or side-chain structural moieties in the peptide. Thirdly, macrocyclization may enhance cell permeability, such as through increased stability of intramolecular hydrogen bonding to reduce the desolvation penalty otherwise incurred in the transport of peptides across an apolar cell membrane. Amongst the several cyclization techniques described, stapling via olefin metathesis using a non-proteogenic amino acid such as alpha-methyl alkenyl side chains has proven to be very effective [13-18], particularly when the desired secondary structure of the peptide macrocycle is helical. Stapling requires incorporation of the appropriate unnatural amino acid precursors to be placed at appropriate locations along the peptide sequence such that they do not interfere with the binding face of the helix. The linkers can be of different types, and can span different lengths, resulting in i,i+3 i,i+4, i,i+7 staples. Although they have largely been used to stabilize helical 3 bioRxiv preprint doi: https://doi.org/10.1101/767673; this version posted September 12, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. conformations, recent studies have also applied RCM strategies to non-helical peptides [19, 20]. The stapled peptide strategy has been successfully applied to inhibit several PPIs of therapeutic potential including, BCL-2/Mcl-1 family [21-24], β-catenin–TCF [25], Rab–GTPase-Effector [26], ERα–coactivator protein [27], Cullin3–BTB [28], VDR–coactivator protein [29], eIf4E [30] and p53–Mdm2/Mdm4 [31-34]. Noteworthy, in the case of p53–Mdm2/Mdm4, a dual selective stapled peptide (ALRN-6924) has been further successfully advanced to phase II clinical trials [35-37]. Although this example is unquestionably encouraging for the advancement of stapled peptides into the clinic, challenges still remain. Amongst these, engineering molecules with sufficient proteolytic stability for sustained target binding and cellular activity is critical. Indeed, although stapling L-amino acid peptides can confer resistance to protease-mediated degradation, the effect is often not complete, and may affect residues located outside of the macrocycle [38-40]. On the other hand, all-D peptides are hyperstable against proteolysis and have been engineered with strong binding affinity against a variety of targets including p53–Mdm2 [41-42], VEGF–VEGF-receptor [43], PD-1–PD-L1 [44], and human immunodeficiency virus type 1 (HIV-1) entry [45]. Unfortunately, although all-D peptides are intrinsically hyperstable to proteolysis, they generally lack membrane permeability and cellular activity. We hypothesized that combining both strategies, ie all-D-peptide and helical macrocyclization might provide synergistic results. Thus, we embarked on introducing a hydrocarbon staple into an all-D peptide inhibitor of the p53 - Mdm2/Mdm4 interaction. p53 is a key tumour suppressor protein, which primarily functions as a DNA transcription factor, which is commonly inactivated in cancer and normally plays a crucial role in guarding the cell in response to various stress signals through the induction of cell cycle arrest, apoptosis or senescence [46]. Mechanisms
Load more

Recommended publications
  • Stapled Α−Helical Peptide Drug Development: a Potent PNAS PLUS Dual Inhibitor of MDM2 and MDMX for P53-Dependent Cancer Therapy
    Stapled α−helical peptide drug development: A potent PNAS PLUS dual inhibitor of MDM2 and MDMX for p53-dependent cancer therapy Yong S. Changa,1,2, Bradford Gravesb,1, Vincent Guerlavaisa, Christian Tovarb, Kathryn Packmanb, Kwong-Him Tob, Karen A. Olsona, Kamala Kesavana, Pranoti Gangurdea, Aditi Mukherjeea, Theresa Bakera, Krzysztof Darlaka, Carl Elkina, Zoran Filipovicb, Farooq Z. Qureshib, Hongliang Caia, Pamela Berryb, Eric Feyfanta, Xiangguo E. Shia, James Horsticka, D. Allen Annisa, Anthony M. Manninga, Nader Fotouhib, Huw Nasha, Lyubomir T. Vassilevb,2, and Tomi K. Sawyera,2 aAileron Therapeutics, Inc., Cambridge, MA 02139; and bRoche Research Center, Hoffmann-La Roche, Inc., Nutley, NJ 07110 Edited* by Robert H. Grubbs, California Institute of Technology, Pasadena, CA, and approved July 12, 2013 (received for review February 17, 2013) Stapled α−helical peptides have emerged as a promising new mo- each unable to compensate for the loss of the other, and they dality for a wide range of therapeutic targets. Here, we report regulate nonoverlapping functions of p53 (4, 6). a potent and selective dual inhibitor of MDM2 and MDMX, The first potent and selective small-molecule inhibitors of the ATSP-7041, which effectively activates the p53 pathway in tumors p53–MDM2 interaction, the Nutlins, provided proof of concept in vitro and in vivo. Specifically, ATSP-7041 binds both MDM2 and that restoration of p53 activity is feasible and may have appli- MDMX with nanomolar affinities, shows submicromolar cellular cation in cancer therapy (11, 12). Although three different activities in cancer cell lines in the presence of serum, and dem- classes of small-molecule MDM2 antagonists are currently under onstrates highly specific, on-target mechanism of action.
    [Show full text]
  • A Short Double-Stapled Peptide Inhibits Respiratory Syncytial Virus Entry and Spreading
    A short double-stapled peptide inhibits respiratory syncytial virus entry and spreading Vanessa Gaillard, Marie Galloux, Dominique Garcin, Jean Francois Eleouet, Ronan Le Goffic, Thibaut Larcher, Marie-Anne Rameix-Welti, Abdelhak Boukadiri, Julien Heritier, Jean-Manuel Segura, et al. To cite this version: Vanessa Gaillard, Marie Galloux, Dominique Garcin, Jean Francois Eleouet, Ronan Le Goffic, et al.. A short double-stapled peptide inhibits respiratory syncytial virus entry and spreading. Antimicrobial Agents and Chemotherapy, American Society for Microbiology, 2017, 61 (4), 10.1128/AAC.02241-16. hal-01605887 HAL Id: hal-01605887 https://hal.archives-ouvertes.fr/hal-01605887 Submitted on 26 May 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution| 4.0 International License ANTIVIRAL AGENTS crossm A Short Double-Stapled Peptide Inhibits Respiratory Syncytial Virus Entry and Spreading Vanessa Gaillard,a Marie Galloux,b Dominique Garcin,c Jean-François Eléouët,b b d e,f Ronan Le Goffic, Thibaut Larcher,
    [Show full text]
  • Stapled Peptides—A Useful Improvement for Peptide-Based Drugs
    molecules Review Stapled Peptides—A Useful Improvement for Peptide-Based Drugs Mattia Moiola, Misal G. Memeo and Paolo Quadrelli * Department of Chemistry, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy; [email protected] (M.M.); [email protected] (M.G.M.) * Correspondence: [email protected]; Tel.: +39-0382-987315 Received: 30 July 2019; Accepted: 1 October 2019; Published: 10 October 2019 Abstract: Peptide-based drugs, despite being relegated as niche pharmaceuticals for years, are now capturing more and more attention from the scientific community. The main problem for these kinds of pharmacological compounds was the low degree of cellular uptake, which relegates the application of peptide-drugs to extracellular targets. In recent years, many new techniques have been developed in order to bypass the intrinsic problem of this kind of pharmaceuticals. One of these features is the use of stapled peptides. Stapled peptides consist of peptide chains that bring an external brace that force the peptide structure into an a-helical one. The cross-link is obtained by the linkage of the side chains of opportune-modified amino acids posed at the right distance inside the peptide chain. In this account, we report the main stapling methodologies currently employed or under development and the synthetic pathways involved in the amino acid modifications. Moreover, we report the results of two comparative studies upon different kinds of stapled-peptides, evaluating the properties given from each typology of staple to the target peptide and discussing the best choices for the use of this feature in peptide-drug synthesis. Keywords: stapled peptide; structurally constrained peptide; cellular uptake; helicity; peptide drugs 1.
    [Show full text]
  • Nonproteinogenic Deep Mutational Scanning of Linear and Cyclic Peptides
    Nonproteinogenic deep mutational scanning of linear and cyclic peptides Joseph M. Rogersa, Toby Passiouraa, and Hiroaki Sugaa,b,1 aDepartment of Chemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan; and bCore Research for Evolutionary Science and Technology, Japan Science and Technology Agency, Saitama 332-0012, Japan Edited by David Baker, University of Washington, Seattle, WA, and approved September 18, 2018 (received for review June 10, 2018) High-resolution structure–activity analysis of polypeptides re- mutants that can be constructed (18). Moreover, it is possible to quires amino acid structures that are not present in the universal combine parallel peptide synthesis with measures of function genetic code. Examination of peptide and protein interactions (19). However, these approaches cannot construct peptide li- with this resolution has been limited by the need to individually braries with the sequence length and numbers that deep muta- synthesize and test peptides containing nonproteinogenic amino tional scanning can, which, at its core, uses high-fidelity nucleic acids. We describe a method to scan entire peptide sequences with acid-directed synthesis of polypeptides by the ribosome. multiple nonproteinogenic amino acids and, in parallel, determine Ribosomal synthesis (i.e., translation) can be manipulated to the thermodynamics of binding to a partner protein. By coupling include nonproteinogenic amino acids (20). In vitro genetic code genetic code reprogramming to deep mutational scanning, any reprogramming is particularly versatile, allowing for the in- number of amino acids can be exhaustively substituted into pep- corporation of amino acids with diverse chemical structures (21). tides, and single experiments can return all free energy changes of Flexizymes, flexible tRNA-acylation ribozymes, can load almost binding.
    [Show full text]
  • Peptide Technologies at Energypolis
    PEPTIDE CHEMISTRY IN SWITZERLAND CHIMIA 2021, 75, No. 6 539 doi:10.2533/chimia.2021.539 Chimia 75 (2021) 539–542 © M. Mathieu, O. Nyanguile Peptide Technologies at Energypolis Marc Mathieu and Origène Nyanguile* Abstract: The new Energypolis campus brings together the skills of EPFL Valais-Wallis, HES-SO Valais-Wallis, and the Ark Foundation’s services. Together these partners respond to today’s major concerns in the domains of energy, health, and the environment cutting-edge technology. The spirit of this new campus is to foster innovation in these disciplines and emulate the creation of start-up companies. The HES-SO hosts the School of Engineering (HEI) at this campus, which includes the following degree programmes: Life Technologies, Systems Engineering and Energy and Environmental Engineering, as well as their corresponding applied research institutes. Peptide technologies belong to the many activities that are carrying out in the Institute of Life Technologies. The present review summarizes the peptide technologies that are currently under development, that is, the regioselective labeling of therapeutic antibodies for cancer imaging, the development of peptide antivirals and antimicrobials for the treatment of infectious diseases, targeting of drugs conjugated to peptidic scaffolds as well as engineer- ing of biomaterials. Keywords: Biomaterials · Cyclic peptides · N0-P complex · Peptide–drug conjugates · Peptidomimetics · Regioselective labeling · Stapled peptides · Therapeutic antibodies 1. Peptides Technologies at Energypolis (Fig. 1) 1.1 Development of a Regioselective Labeling Method for Clinical Imaging of Therapeutic Antibodies In a collaboration between Debiopharm Research & Manufacturing, the CHUV, and the HES-SO Valais-Wallis, a novel methodology was developed to regioselectively label therapeutic antibodies with an imaging cargo.
    [Show full text]
  • Functionalised Staple Linkages for Modulating The
    Chemical Science Accepted Manuscript This is an Accepted Manuscript, which has been through the Royal Society of Chemistry peer review process and has been accepted for publication. Accepted Manuscripts are published online shortly after acceptance, before technical editing, formatting and proof reading. Using this free service, authors can make their results available to the community, in citable form, before we publish the edited article. We will replace this Accepted Manuscript with the edited and formatted Advance Article as soon as it is available. You can find more information about Accepted Manuscripts in the Information for Authors. Please note that technical editing may introduce minor changes to the text and/or graphics, which may alter content. The journal’s standard Terms & Conditions and the Ethical guidelines still apply. In no event shall the Royal Society of Chemistry be held responsible for any errors or omissions in this Accepted Manuscript or any consequences arising from the use of any information it contains. www.rsc.org/chemicalscience Page 1 of 6 Chemical Science Chemical Science RSCPublishing EDGE ARTICLE Functionalised staple linkages for modulating the cellular activity of stapled peptides Cite this: DOI: 10.1039/x0xx00000x Yu Heng Lau,a Peterson de Andrade,a Soo-Tng Quah,b Maxim Rossmann,c Luca Laraia,a,f Niklas Sköld,a Tze Jing Sum,a Pamela J. E. Rowling,d Thomas L. e e c d Received 00th January 2012, Joseph, Chandra Verma, Marko Hyvonen, Laura S. Itzhaki, Ashok R. f b b a* Accepted 00th January 2012 Venkitaraman, Christopher J. Brown, David P. Lane and David R.
    [Show full text]
  • Rapid Optimization of Mcl-1 Inhibitors Using Stapled Peptide Libraries Including Non-Natural Side Chains
    Rapid Optimization of Mcl-1 Inhibitors using Stapled Peptide Libraries Including Non-Natural Side Chains The MIT Faculty has made this article openly available. Please share how this access benefits you. Your story matters. Citation Rezaei Araghi, Raheleh et al. “Rapid Optimization of Mcl-1 Inhibitors Using Stapled Peptide Libraries Including Non-Natural Side Chains.” ACS Chemical Biology 11, 5 (February 2016): 1238–1244 © 2016 American Chemical Society As Published http://dx.doi.org/10.1021/ACSCHEMBIO.5B01002 Publisher American Chemical Society (ACS) Version Author's final manuscript Citable link http://hdl.handle.net/1721.1/116641 Terms of Use Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. HHS Public Access Author manuscript Author ManuscriptAuthor Manuscript Author ACS Chem Manuscript Author Biol. Author Manuscript Author manuscript; available in PMC 2017 May 20. Published in final edited form as: ACS Chem Biol. 2016 May 20; 11(5): 1238–1244. doi:10.1021/acschembio.5b01002. Rapid Optimization of Mcl-1 Inhibitors using Stapled Peptide Libraries Including Non-Natural Side Chains Raheleh Rezaei Araghi1, Jeremy A. Ryan2, Anthony Letai2,3, and Amy E. Keating1,4,* 1Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Ave. Cambridge MA 02139, United States 2Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, United States 3Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, United States 4Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave. Cambridge MA 02139, United States Abstract Alpha helices form a critical part of the binding interface for many protein-protein interactions, and chemically stabilized synthetic helical peptides can be effective inhibitors of such helix- mediated complexes.
    [Show full text]
  • Investigation of Nitrile Oxide-Alkyne 1,3-Dipolar Cycloadditions and Their Potential Viability for Synthesis of Stapled Peptides
    Investigation of Nitrile Oxide-Alkyne 1,3-Dipolar Cycloadditions and Their Potential Viability for Synthesis of Stapled Peptides by Charles Teeples A thesis presented to the Honors College of Middle Tennessee State University in partial fulfillment of the requirements for graduation from the University Honors College. Fall 2020 Investigation of Nitrile Oxide-Alkyne Cycloadditions and Their Potential Viability for Synthesis of Stapled Peptides by Charles Teeples APPROVED: ______________________________________ Dr. Norma Dunlap, Thesis Director Professor, Department of Chemistry ______________________________________ Dr. Scott Handy, Second Reader Professor, Department of Chemistry ______________________________________ Dr. Gregory Van Patten, Thesis Committee Chair Chair, Department of Chemistry ii Acknowledgments This project would not be possible without my advisor, Dr. Norma Dunlap. I appreciate all the time she spent teaching me the basics of organic synthesis in the lab, as well as the writing advice she gave me outside the laB. I also want to thank my mom and grandmother (Jane and Jewell) who supported me and encouraged my academic pursuits. Finally, I want to thank my girlfriend, Ida, who gave me support during the long days and nights needed to finish this project. iii Abstract The nitrile oxide-alkyne 1,3-dipolar cycloaddition reaction is a concerted reaction between a nitrile oxide, which is usually generated in situ, and an alkyne to afford an isoxazole. This reaction has been previously used in post-synthetic modification of nucleic acids. This work examines the potential utility of the nitrile oxide-alkyne 1,3- dipolar cycloaddition reaction in the synthesis of stapled peptides. Stapled peptides, or hydrocarbon cross-linked peptides, show potential as drugs to target protein-protein interactions in the body.
    [Show full text]
  • 1 Design and Development of Stapled Transmembrane Peptides That
    Design and development of stapled transmembrane peptides that disrupt the activity of G-protein coupled receptor oligomers Joaquín Botta1,2, Lucka Bibic2, Patrick Killoran3, Peter J. McCormick*1 and Lesley A. Howell*4 From the 1Centre for Endocrinology, William Harvey Research Institute, Bart’s and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London, EC1M 6BQ; 2School of Pharmacy, University of East Anglia, Norwich Research Park, Norwich, NR4 7TJ; 3School of Pharmacy and Biomolecular Sciences, James Parsons Building, Byrom Street, Liverpool, L3 3AF; 4School of Biological and Chemical Sciences, Queen Mary University of London, Mile End Road, London, E1 4NS Running Title: Stapled TM peptides modulate GPCR oligomers * To whom correspondence should be addressed: Lesley A. Howell: School of Biological and Chemical Sciences, Queen Mary University of London, Mile End Road, London, E1 4NS; [email protected]; Tel. (+44) 207 8826625 or Peter J. McCormick: Centre for Endocrinology, William Harvey Research Institute, Bart’s and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London, EC1M 6BQ; [email protected]. Keywords: G protein‐coupled receptor (GPCR), cannabinoid receptor type 1 (CB1), dimerization, cell‐ penetrating peptide (CPP), peptide chemical synthesis, serotonin receptor type 2A (5HT2A), NanoLuc binary technology (NanoBiT), hydrocarbon stapling, transmembrane peptide, bimolecular fluorescence complementation assessments, we developed a NanoLuc binary ABSTRACT technology (NanoBiT)-based reporter assay to Membrane proteins can associate into screen a small library of aryl-carbon–stapled larger complexes. Examples include receptor transmembrane mimicking peptides produced by tyrosine complexes, ion channels, transporters solid-phase peptide synthesis.
    [Show full text]
  • Peptide Stapling Techniques Based on Different Macrocyclisation Chemistries
    Chemical Society Reviews Peptide stapling techniques based on different macrocyclisation chemistries Journal: Chemical Society Reviews Manuscript ID: CS-TRV-07-2014-000246.R1 Article Type: Tutorial Review Date Submitted by the Author: 16-Aug-2014 Complete List of Authors: Lau, Yu Heng; University of Cambridge, Chemistry de-Andrade, Peterson; University of Cambridge, Chemistry Wu, Yuteng; University of Cambridge, Chemistry Spring, David; University of Cambridge, Department of Chemistry Page 1 of 12 Chemical Society Reviews Chemical Society Reviews RSC Publishing TUTORIAL REVIEW Peptide stapling techniques based on different macrocyclisation chemistries Cite this: DOI: 10.1039/x0xx00000x Yu Heng Lau, Peterson de Andrade, Yuteng Wu and David R. Spring* Received 00th January 2012, Peptide stapling is a strategy for constraining short peptides typically in an alpha-helical Accepted 00th January 2012 conformation. Stapling is carried out by covalently linking the side-chains of two amino acids, thereby forming a peptide macrocycle. There is an expanding repertoire of stapling techniques DOI: 10.1039/x0xx00000x based on different macrocyclisation chemistries. In this tutorial review , we categorise and www.rsc.org/ analyse key examples of peptide stapling in terms of their synthesis and applicability to biological systems. Key Learning Points 1. A growing number of stapling techniques are being developed, each utilising different macrocyclisation chemistry. 2. Stapling techniques can be broadly categorised as one- or two-component reactions. 3. Different stapling chemistries have strengths and weaknesses in terms of both their synthetic ease and potential utility for addressing biological questions. 4. Two-component reactions enable efficient modular construction of peptides bearing different staple linkages, orthogonal to changes in the peptide sequence.
    [Show full text]
  • Stapled Peptide Targeting the CDK4/Cyclin D Interface Combined
    Stapled peptide targeting the CDK4/Cyclin D interface combined with Abemaciclib inhibits KRAS mutant lung cancer growth Céline Bouclier, Matthieu Simon, Guillaume Laconde, Morgan Pellerano, Sebastien Diot, Sylvie Lantuejoul, Benoit Busser, Laetitia Vanwonterghem, Julien Vollaire, Véronique Josserand, et al. To cite this version: Céline Bouclier, Matthieu Simon, Guillaume Laconde, Morgan Pellerano, Sebastien Diot, et al.. Sta- pled peptide targeting the CDK4/Cyclin D interface combined with Abemaciclib inhibits KRAS mu- tant lung cancer growth. Theranostics, Ivyspring International Publisher, 2020, 10 (5), pp.2008-2028. 10.7150/thno.40971. inserm-02440284 HAL Id: inserm-02440284 https://www.hal.inserm.fr/inserm-02440284 Submitted on 15 Jan 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Theranostics 2020, Vol. 10, Issue 5 2008 Ivyspring International Publisher Theranostics 2020; 10(5): 2008-2028. doi: 10.7150/thno.40971 Research Paper Stapled peptide targeting the CDK4/Cyclin D interface combined with Abemaciclib inhibits KRAS mutant lung cancer growth Celine Bouclier1,*, Matthieu Simon1,*, Guillaume Laconde1, Morgan Pellerano1, Sebastien Diot1, Sylvie Lantuejoul2, Benoit Busser2,3, Laetitia Vanwonterghem3, Julien Vollaire3, Véronique Josserand3, Baptiste Legrand1, Jean-Luc Coll3, Muriel Amblard1, Amandine Hurbin3,#, May C.
    [Show full text]
  • Unprotected Peptide Macrocyclization and Stapling Via a Fluorine-Thiol
    bioRxiv preprint doi: https://doi.org/10.1101/2020.09.09.290379; this version posted September 9, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Unprotected Peptide Macrocyclization and Stapling via A Fluorine-Thiol Displacement Reaction Md Shafiqul Islam1, Samuel L. Junod2, Si Zhang1, Zakey Yusuf Buuh1, Yifu Guan1, Kishan H Kaneria1, Zhigang Lyu1, Vincent Voelz1, Weidong Yang2, Rongsheng E. Wang1,* 1. Department of Chemistry, Temple University, 1901 N. 13th Street, Philadelphia, PA, 19122 2. Department of Biology, Temple University, 1900 N. 12th Street, Philadelphia, PA, 19122 * To whom correspondence should be addressed: Phone (215) 204-1855. Email: [email protected] 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.09.09.290379; this version posted September 9, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract Stapled peptides serve as a powerful tool for probing protein-protein interactions, but its application has been largely impeded by the limited cellular uptake. Here we report the discovery of a facile peptide macrocyclization and stapling strategy based on a fluorine thiol displacement reaction (FTDR), which renders a class of peptide analogues with enhanced stability, affinity, and cell permeability. This new approach enabled selective modification of the orthogonal fluoroacetamide side chains in unprotected peptides, with the identified 1,3- benzenedimethanethiol linker promoting alpha helicity of a variety of peptide substrates, as corroborated by molecular dynamics simulations.
    [Show full text]